An improved HPLC method with fluorescence detection for the determination of pyrene in rat plasma and its pharmacokinetics.
A high-performance liquid chromatographic method with fluorescence detection (HPLC-FLD) for the determination of pyrene in rat plasma was developed and validated. The method used fluorene as internal standard (IS), following a single-step protein precipitation, the analyte and internal standard were separated on a C18 column with a mobile phase containing methanol-water (90:10, v/v) at a flow rate of 1 ml/min. The analytes were detected by using fluorescence detection at an excitation and emission wavelength of 265 and 394 nm, respectively. Two calibration curves were constructed in the range of 2-100 ng/ml and 0.1-5 microg/ml for pyrene with a lower limit of quantitation (LLOQ) of 2 ng/ml. Both intra-day and inter-day precision were less than 6% except at LLOQ, for which the precision was 10.6 and 9.8, respectively. Accuracy ranged from 98.3 to 103.6%, except at LLOQ, for which the accuracy was about 85%. The recovery ranged from 84.7 to 95.0% at the low, medium and high concentrations. The present HPLC-FLD method was rapid, sensitive, and reliable. The method described herein had been successfully applied for the pharmacokinetic studies in female Wistar rats after administration of 10mg equivalent pyrene/kg dose of solution of pyrene and 1mg equivalent pyrene/kg dose of pyrene-loaded nanoparticle.
Zhao
Xiaozhi
X
College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, PR China.
Wan
Jiangling
J
Xu
Huibi
H
Yang
Xiangliang
X
D000818:Animals; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004355:Drug Stability; D005260:Female; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry / Q000493:pharmacokinetics*; D005615:Freezing; D015394:Molecular Structure; D053758:Nanoparticles; D010316:Particle Size; D011721:Pyrenes / Q000032:analysis* / Q000737:chemistry / Q000493:pharmacokinetics*; D051381:Rats; D017208:Rats, Wistar; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000379:methods; D013997:Time Factors
D005456:Fluorescent Dyes; D011721:Pyrenes; C030984:pyrene